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Journal of Virology, October 2006, p. 9608-9618, Vol. 80, No. 19
0022-538X/06/$08.00+0     doi:10.1128/JVI.00850-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.

Adenovirus Core Protein pVII Is Translocated into the Nucleus by Multiple Import Receptor Pathways{dagger}

Harald Wodrich,1,3*,{ddagger} Aurelia Cassany,1,{ddagger} Maximiliano A. D'Angelo,1,§ Tinglu Guan,1 Glen Nemerow,2 and Larry Gerace1

Department of Cell Biology, Scripps Research Institute, La Jolla, California,1 Department of Immunology, Scripps Research Institute, La Jolla, California,2 Institut de Génétique Moléculaire de Montpellier, UMR 5535 CNRS, IFR 122, Montpellier, France3

Received 25 April 2006/ Accepted 6 July 2006

Adenoviruses are nonenveloped viruses with an ~36-kb double-stranded DNA genome that replicate in the nucleus. Protein VII, an abundant structural component of the adenovirus core that is strongly associated with adenovirus DNA, is imported into the nucleus contemporaneously with the adenovirus genome shortly after virus infection and may promote DNA import. In this study, we evaluated whether protein VII uses specific receptor-mediated mechanisms for import into the nucleus. We found that it contains potent nuclear localization signal (NLS) activity by transfection of cultured cells with protein VII fusion constructs and by microinjection of cells with recombinant protein VII fusions. We identified three NLS-containing regions in protein VII by deletion mapping and determined important NLS residues by site-specific mutagenesis. We found that recombinant protein VII and its NLS-containing domains strongly and specifically bind to importin {alpha}, importin ß, importin 7, and transportin, which are among the most abundant cellular nuclear import receptors. Moreover, these receptors can mediate the nuclear import of protein VII fusions in vitro in permeabilized cells. Considered together, these data support the hypothesis that protein VII is a major NLS-containing adaptor for receptor-mediated import of adenovirus DNA and that multiple import pathways are utilized to promote efficient nuclear entry of the viral genome.

* Corresponding author. Mailing address: Institut de Génétique Moléculaire de Montpellier, UMR 5535 CNRS, 1919 Route de Mende, 34293 Montpellier Cedex 05, France. Phone: 33 4 67 61 36 74. Fax: 33 4 67 04 02 31. E-mail: harald.wodrich{at}igmm.cnrs.fr.

{dagger} Supplemental material for this article may be found at http://jvi.asm.org.

{ddagger} H.W. and A.C. contributed equally to this work.

§ Present address: Molecular and Cell Biology Laboratory, Salk Institute for Biological Studies, La Jolla, Calif.

Journal of Virology, October 2006, p. 9608-9618, Vol. 80, No. 19
0022-538X/06/$08.00+0     doi:10.1128/JVI.00850-06
Copyright © 2006, American Society for Microbiology. All Rights Reserved.



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